754 .
MJ MEDIUM FOR DEFFERIBACTER SP. STRAIN NEJINEJI
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 30.00 | g | ||
|
|
MgCl2 x 6 H2O | 6.50 | g | ||
|
|
CaCl2 x 2 H2O | 0.50 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
K2HPO4 | 0.14 | g | ||
|
|
MnCl2 x 4 H2O | 4.00 | mg | ||
|
|
Na2SeO3 x 5 H2O | 4.00 | mg | ||
|
|
NiCl2 x 6 H2O | 2.00 | mg | ||
|
|
CoCl2 x 6 H2O | 2.00 | mg | ||
|
|
ZnCl2 | 0.50 | mg | ||
|
|
Na2MoO4 x 2 H2O | 0.40 | mg | ||
|
|
FeCl2 x 4 H2O | 0.25 | mg | ||
|
|
Na2WO2 x 2 H2O | 0.10 | mg | ||
|
|
H3BO3 | 0.10 | mg | ||
|
|
Sulfur | 10.00 | g | ||
|
|
Resazurin | 0.50 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components except sulfur and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions from anaerobic stocks (filter-sterilized): | |||||
|
|
Trace vitamins | 1.00 | ml | ||
|
|
NaHCO3
(8%) |
12.50 | ml | ||
| 2 Separately steam sulfur for 3 hr on each of 3 successive days. | |||||
| 3 To complete the medium, distribute the solution into sterile culture vessels containing appropriate amount of sulfur (e.g., 5 ml solution and 0.05 g sulfur in Hungate tubes) under a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Then, add Na2S x 9H2O solution from an anaerobic stock (final 0.1 g/L). After inoculation, pressurize the inoculated culture vessels to 200 kPa H2-CO2 (4:1, v/v). | |||||
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |